BackgroundCAR T cell therapy is remarkably successful in patients with hematological malignancies, in some cases inducing durable remissions. However, it remains ineffective in solid tumors, in part ...due to poor T cell infiltration into the tumor mass. Determinants of successful T cell infiltration to the tumor site remain to be defined. In contrast, tumors actively attract T regulatory (Treg) cells for immune suppression through the C-C chemokine receptor 8 (CCR8) - CCL1 axis. As this axis is functional across cancer entities, we postulated that CCR8 could also be used to target tumor-ablating T cells to the tumor site.Material and methodsMurine and human CCR8 have been cloned in a retroviral expression vector. CCR8 can be expressed in murine and human T cells upon transduction. A chimeric antigen receptor (CAR) targeting the murine epithelial cell adhesion molecule (EpCAM) was used for syngeneic pancreatic tumor models and a CAR targeting human mesothelin was used for a xenograft pancreatic tumor model.Mechanistically, we use flow cytometry and multi-photon intra-vital microscopy to interrogate infiltration of CCR8-transduced CAR T cells.ResultsHere we show that genetically engineering CAR T cells to express CCR8 improves their migration into solid tumors and allows rejection of tumors that are otherwise resistant to CAR T cell therapy. We demonstrate the capacity of these enhanced CAR T cells to stunt solid tumor growth and improve survival in both murine syngeneic and human xenograft tumor models.ConclusionOur results demonstrate the viability of using CCR8 to confer Tregcell trafficking-properties in CAR T cells to enable their effectiveness in solid tumors. This receptor may be combined with other promising strategies to improve the efficacy of cellular approaches.Disclosure InformationB. L. Cadilha: None. K. Dorman: None. D. Huynh: None. T. Lorenzini: None. M. Vänttinen: None. M. -R. Benmebarek: None. S. Stoiber: None. J. Suárez-Gosálvez: None. S. Endres: None. S. Kobold: None.
Adoptive T cell therapy (ACT) is a therapeutic approach which employs genetically manipulated autologous T cells to target and eliminate a patient's malignancy. This novel therapeutic approach, when ...employing a chimeric antigen receptor (CAR) targeting CD19-expressing B cells, has shown remarkable success in treating acute B-cell lymphocytic leukemia. However, blood born malignancies represent only a fraction of cancers which affect patients. Unfortunately, the utilization of ACT to target solid malignancies has only shown marginal success rates. There are many known obstacles which hinder CAR T cell therapy in patients suffering from solid cancer, one notable obstacle is the effective trafficking of CAR T cells to the tumor site. With the rapid advancement of novel approaches and targets which may enhance CAR T cell infiltration into solid tumors, a standardized approach to assess and measure CAR T cell infiltration becomes imperative in order to compare these different approaches across platforms. Here we describe a flow cytometry method which enables the rapid detection and quantification of CAR T cells which have reached and entered the tumor mass following intravenous injection. Competence with single cell preparation and flow cytometry is required for optimal results.
During metastasis, cancer cells invade, intravasate, enter the circulation, extravasate, and colonize target organs. Here, we examined the role of interleukin (IL)-22 in metastasis. Immune ...cell-derived IL-22 acts on epithelial tissues, promoting regeneration and healing upon tissue damage, but it is also associated with malignancy. Il22-deficient mice and mice treated with an IL-22 antibody were protected from colon-cancer-derived liver and lung metastasis formation, while overexpression of IL-22 promoted metastasis. Mechanistically, IL-22 acted on endothelial cells, promoting endothelial permeability and cancer cell transmigration via induction of endothelial aminopeptidase N. Multi-parameter flow cytometry and single-cell sequencing of immune cells isolated during cancer cell extravasation into the liver revealed iNKT17 cells as source of IL-22. iNKT-cell-deficient mice exhibited reduced metastases, which was reversed by injection of wild type, but not Il22-deficient, invariant natural killer T (iNKT) cells. IL-22-producing iNKT cells promoting metastasis were tissue resident, as demonstrated by parabiosis. Thus, IL-22 may present a therapeutic target for prevention of metastasis.
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•Il22-deficient mice are protected against metastasis formation•IL-22 neutralization blocks cancer cell extravasation•IL-22 acts on endothelial cells, promoting cancer cell extravasation via ANPEP induction•Tissue resident iNKT17 cells are the key IL-22 source during cancer cell extravasation
Interleukin-22 (IL-22) is produced by immune cells and promotes tissue repair and regeneration; however, in malignancy, IL-22 can promote tumor growth. Giannou et al. find that tissue resident iNKT17 cells produce IL-22 and promote cancer cell extravasation through regulation of aminopeptidase N. Neutralization of IL-22 inhibits metastasis formation, suggesting therapeutic avenues for cancer treatment.
BackgroundHigh expression of CD155 (poliovirus receptor, PVR) is associated with a poor prognosis of lung adenocarcinoma (LUAD) and triple-negative breast cancer (TNBC) patients. When overexpressed, ...this molecule inhibits the antitumor function of NK and cytotoxic T cells through binding to its inhibitory co-receptors TIGIT and CD96, and downregulation of stimulatory CD226 (DNAM-1). However, the exact mechanism of CD155 overexpression on the tumor cells remains unclear. Here we demonstrate that interleukin-22 (IL-22), a cytokine known to promote cancer progression, induces upregulation of CD155 on tumor cells in mouse models of breast and lung cancer and may, thus, inhibit antitumor immunity and promote lung metastasis.Materials and MethodsTo study the influence of IL-22 on antitumor immunity, we utilize IL-22-deficient animals in syngeneic mouse models of metastatic breast and lung cancer. For this purpose, we generated tumor cells deficient in IL-22 receptor (IL-22R) or in CD155 and tumor cells, that constantly express CD155 independent of its natural regulation. Here, we determine the incidence of metastasis and antitumor NK and T cell responses in the lung, the primary site of metastasis.ResultsWe demonstrate that murine cancer cells upregulate CD155 surface expression upon treatment with recombinant IL-22, whereas this effect is abolished in the absence of IL-22R. Furthermore, IL-22-deficient animals have a lower metastatic burden in the lung and demonstrate a dramatic increase in IFN-γ production in NK, and, to a lower extent, cytotoxic T cells. Moreover, this effect is reversed when CD155 is expressed on the tumor cells independent of its natural regulation, which enables lung metastases in IL-22 deficient animals. Phenotypically, NK cells in IL-22 knockout mice have a higher expression of co-stimulatory receptor CD226, which is linked to the antitumor potential of these cells.ConclusionsHere we demonstrate a novel pathway of cytokine-mediated cancer progression, where IL-22 is capable of inducing CD155 on the tumor cells and, therefore, promotes an immunosuppressive tumor microenvironment. This highlights the potential of IL-22 as a target for immunotherapy considering the complexity of the CD155-dependent immunoregulatory network.Disclosure InformationD. Briukhovetska: None. J. Suarez-Gosalvez: None. M. Schübel: None. A. Markota: None. J. Jobst: None. J. Dörr: None. F. Märkl: None. M. Schwerdtfeger: None. A. Öner: None. M. Seifert: None. A. Gottschlich: None. S. Endres: None. S. Kobold: None.
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